Bacterial diversity of two distinct wastewater treatment systems, conventional activated sludge (CAS) and membrane bioreactor (MBR), of petroleum refineries were investigated through 16S rRNA gene libraries. Sequencing and phylogenetic analysis showed that the bacterial community composition of sludge samples was distinct between the two wastewater treatment systems. MBR clones belonged predominantly to Class Betaproteobacteria, represented mainly by genera Thiobacillus and Thauera, whereas CAS clones were mostly related to Class Alphaproteobacteria, represented by uncultured bacteria related to Order Parvularculales. Richness estimators ACE and Chao revealed that the diversity observed in both libraries at the species level is an underestimate of the total bacterial diversity present in the environment and further sampling would yield an increased observed diversity. Shannon and Simpson diversity indices were different between the libraries and revealed greater bacterial diversity for the MBR library, considering an evolutionary distance of 0.03. LIBSHUFF analyses revealed that MBR and CAS communities were significantly different at the 95% confidence level () for distances . This work described, qualitatively and quantitatively, the structure of bacterial communities in industrial-scale MBR and CAS processes of the wastewater treatment system from petroleum refineries and demonstrated clearly differentiated communities responsible for the stable performance of wastewater treatment plants.

A mutant of green fluorescent protein () from the jellyfish Aequorea victoria was cyclized in vitro and in vivo by the use of a naturally split intein from the dnaE gene of Synechocystis species PCC6803 (Ssp). Cyclization of was confirmed by amino acid sequencing and resulted in an increased electrophoretic mobility compared with the linear . The circular was more thermostable than the linear form and significantly more resistant to proteolysis of exopeptidase. The circular also displayed increased relative fluorescence intensity. In addition, chemical stability of against GdnHCl revealed more stability of the circular form compared with the linear form.

To improve the expression efficiency of recombinant endo--1,4-glucanase in P. pastoris, the endo--1,4-glucanase (egI) gene from Aspergillus niger was synthesized using optimized codons. Fourteen pairs of oligonucleotides with 15 bp overlap were designed and the full-length syn-egI gene was generated by two-step PCR-based DNA synthesis. In the synthesized endo--1,4-glucanase gene syn-egI, 193 nucleotides were changed, and the G+C content was decreased from 54% to 44.2%. The syn-egI gene was inserted into pPIC9K and transformed into P. pastoris GS115 by electroporation. The enzyme activity of recombinant P. pastoris stain 2-7# reached 20.3 U/ml with 1% barley -glucan and 3.3 U/ml with 1% carboxymethylcellulose (CMC) as substrates in shake flasks versus 1,270.3 U/ml and 220.7 U/ml for the same substrates in 50-1 fermentors. The molecular mass of the recombinant protein was approximately 40 kDa as determined by SDS-PAGE analysis, the optimal temperature for recombinant enzyme activity was , and the optimal pH was 5.0 when CMC was used as the substrate.

In this paper, we analyzed the microsatellites in the transcript sequences of the whole Laccaria bicolor genome. Our results revealed that, apart from the triplet repeats, length diversification and richness of the detected microsatellites positively correlated with their repeat motif lengths, which were distinct from the variation trends observed for the transcriptional microsatellites in the genome of higher plants. We also compared the microsatellites detected in the genic regions and in the nongenic regions of the L. bicolor genome. Subsequently, SSR primers were designed for the transcriptional microsatellites in the L. bicolor genome. These SSR primers provide desirable genetic resources to the ectomycorrhizae community, and this study provides deep insight into the characteristics of the micro satellite sequences in the L. bicolor genome.

AfsR2 is a global regulatory protein that stimulates antibiotic biosynthesis in both Streptomyces lividans and S. coelicolor. Previously, various afsR2-dependent genes including a putative abaA-like regulatory gene, SCO4677, were identified through comparative DNA microarray analysis. To further identify the putative SCO4677-dependent proteins, the comparative proteomics-driven approach was applied to the SCO4677-overexpressing strains of S. lividans and S. coelicolor along with the wild-type strains. The 2D gel electrophoresis gave approximately 277 protein spots for S. lividans and 207 protein spots for S. coelicolor, showing different protein expression patterns between the SCO4677-overexpressing strains and the wild-type strains. Further MALDI-TOF analysis revealed that only 18 proteins exhibited similar expression patterns in both S. lividans and S. coelicolor, suggesting that the SCO4677 could encode an abaA-like regulator that controls a few cross-species common proteins as well as many species-specific proteins in Streptomyces species.

A modified graphite felt electrode with neutral red (NR-electrode) was shown to catalyze the chemical oxidation of nitrite to nitrate under aerobic conditions. The electrochemically oxidized NR-electrode (EO-NR-electrode) and reduced NR-electrode (ER-NR-electrode) catalyzed the oxidation of mg/l and mg/l of nitrite, respectively, for 24 h. The electrically uncharged NR-electrode (EU-NR-electrode) catalyzed the oxidation of mg/l of nitrite for 24 h. The aerobic bacterial community immobilized in the EO-NR-electrode did not oxidize ammonium to nitrite; however, the aerobic bacterial community immobilized in the ER-NR-electrode bioelectrochemically oxidized mg/l of ammonium for 48 h. Meanwhile, the aerobic bacterial community immobilized on the EU-NR-electrode biochemically oxidized mg/l of ammonium for 48 h. In the continuous culture system, the aerobic bacterial community immobilized on the ER-NR-electrode bioelectrochemically oxidized a minimal mg/l to a maximal mg/l of ammonium to nitrate, and the community immobilized on the EU-NR-electrode biochemically oxidized a minimal mg/l to a maximal mg/l of ammonium to nitrate every two days. The bacterial communities cultivated in the ER-NR-electrode and EU-NR-electrode in the continuous culture system were analyzed by TGGE on the and days of incubation. Some ammonium-oxidizing bacteria were enriched on the ER-NR-electrode, but not on the EU-NR-electrode.

Bacterial blight, an important and potentially destructive bacterial disease in rice, is caused by Xanthomonas oryzae. Recently, this organism has developed resistance to available antibiotics, prompting scientists to find a suitable alternative. This study focused on secondary metabolites of Phomopsis longicolla to target X. oryzae. Five bioactive compounds were isolated by activity-guided fractionation from ethyl acetate extracts of mycelia and were identified by LC/MS and NMR spectroscopy as dicerandrol A, dicerandrol B, dicerandrol C, deacetylphomoxanthone B, and fusaristatin A. This is the first time fusaristatin A has been isolated from Phomopsis sp. Deacetylphomoxanthone B showed a higher antibacterial effect against X. oryzae KACC 10331 than the positive control (2,4-diacetyphloroglucinol). Dicerandrol A also showed high antimicrobial activity against Gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis) and yeast (Candida albicans). In addition, high production yields of these compounds were obtained at the stationary and death phases.

1-Acetyl--carboline was isolated as an anti-MRSA agent from the fermentation broth of a marine actinomycete isolated from marine sediment. The producing strain was identified to be Streptomyces sp. by phylogenetic analysis of the 16S rRNA gene sequence. The anti-MRSA agent was isolated by bioactivity-guided fractionation of the culture extract by solvent partitioning, ODS open flash chromatography, and purification with a reversed-phase HPLC. Its structure was elucidated by extensive 2D NMR and mass spectral analyses. Combination of 1-acetyl--carboline with ampicillin exhibited synergistic antibacterial activity against MRSA.

The application of Beauveria bassiana SFB-205 supernatant incorporated with polyoxyethylene-(3)-isotridecyl ether (TDE-3) significantly reduced the population of two species of aphids including cotton aphid, Aphis gossypii, and green peach aphid, Myzus persicae, much higher in cotton aphid, compared with supernatant incorporated with Tween 80, which allows the relationship of aphicidal activity with the degradation of aphid cuticles to be determined. Overall, the degradation of the cuticles induced by the supernatant was more remarkable in conjunction with TDE-3 than Tween 80, and this phenomenon was more observable in cotton aphid through SDS-polyacrylamide gel electrophoresis, revealing high correlation with their aphicidal activities.

A series of thiobarbituric acid derivatives were constructed and evaluated for inhibitory activity on hepatitis C virus NS5B polymerase. In biochemical assays using purified viral polymerase and RNA template, the value was improved to 0.41 from the original compound's 1.7 value. In HCV sub genomic replicon assay, the value was improved to 3.7 from the original compound's 12.3 value. was higher than 77 for all compounds tested, suggesting that they are useful candidates for anti-HCV therapy.

A thermostable trehalose synthase (TtTSase) from Thermus thermophilus HJ6 was immobilized on chitosan activated with glutaraldehyde. The yield of immobilization was evaluated as 39.68%. The optimum pH of the immobilized enzyme was similar to that of the free enzyme. However, the optimal temperature ranges were shifted by about owing to better thermal stability after immobilization. The half-life of heat inactivation for free and immobilized enzymes was 5.7 and 6.3 days at , respectively, thus showing a lager thermostability of the immobilized enzyme. When tested in batch reaction, the immobilized enzyme retained its relative activity of 53% after 30 reuses of reaction within 12 days, and still retained 82% of its initial activity even after 150 days at . A packed-bed bioreactor with immobilized enzyme showed a maximum yield of 56% trehalose from 100 mM maltose in a continuous recycling system (bed volume: 10 ml) under conditions of pH 7.0 and .

A mannanase gene (man26B) was obtained from a sea bacterium, Paenibacillus sp. BME-14, through the constructed genomic library and inverse PCR. The gene of man26B had an open reading frame of 1,428 bp that encoded a peptide of 475- amino acid residues with a calculated molecular mass of 53 kDa. Man26B possessed two domains, a carbohydrate binding module (CBM) belonging to family 6 and a family 26 catalytic domain (CD) of glycosyl hydrolases, which showed the highest homology to Cel44C of P. polymyxa (60% identity). The optimum pH and temperature for enzymatic activity of Man26B were 4.5 and , respectively. The activity of Man26B was not affected by and , but was inhibited by , and -mercaptoethanol, and slightly enhanced by and . EDTA did not affect the activity of Man26B, which indicates that it does not require divalent ions to function. Man26B showed a high specific activity for LBG and konjac glucomannan, with , and values of 3.80 mg/ml, 91.70 /min/mg protein, and 77.08/s, respectively, being observed when LBG was the substrate. Furthermore, deletion of the CBM6 domain increased the enzyme stability while enabling it to retain 80% and 60% of its initial activity after treatment at and for 30 min, respectively. This finding will be useful in industrial applications of Man26B, because of the harsh circumstances associated with such processes.

This work was aimed at utilizing rice bran as a substrate for -carotene production by Rhodotorula glutinis DM 28 under optimized conditions of solid-state fermentation. The biomass and -carotene content of Rhodotorula glutinis DM 28 grown on rice bran as a sole substrate under solid-state fermentation were 54 g/kg rice bran and 1.65 mg/kg rice bran, respectively. Its biomass and -carotene content, however, could be improved by 60% and 30%, respectively, using the Central Composite Design for the optimization of its cultivation conditions. The optimized conditions obtained were a pH of 5, a moisture content of 70% (w/w), and a carbon-to-nitrogen ratio of 4. Under these conditions, rice bran containing R. glutinis DM 28 had nutritional values of -carotene, protein, and fat higher than those of rice bran alone. Yeast-grown rice bran could be suitable, therefore, to use as a -carotene-enriched supplement in animal feeds.

Peroxidase-like activity of Vitreoscilla hemoglobin (VHb) has been recently disclosed. To maximize such activity, two catalytically conserved residues (histidine and arginine) found in the distal pocket of peroxidases have successfully been introduced into that of the VHb. A 15-fold increase in catalytic constant () was obtained in P54R variant,which was presumably attributable to the lower rigidity and higher hydrophilicity of the distal cavity arising from substitution of proline to arginine. None of the modifications altered the affinity towards either or ABTS substrate. Spectroscopic studies revealed that VHb variants harboring the T29H mutation apparently demonstrated a spectral shift in both ferric and ferrous forms (406-408 to 411 nm, and 432 to 424-425 nm, respectively). All VHb proteins in the ferrous state had a peak at ~419 nm following the carbon monoxide (CO) binding. Expression of the P54R mutant mediated the downregulation of iron superoxide dismutase (FeSOD) as identified by two-dimensional gel electrophoresis (2-DE) and peptide mass fingerprinting (PMF). According to the high peroxidase activity of P54R, it could effectively eliminate autoxidation-derived , which is a cause of heme degradation and iron release. This decreased the iron availability and consequently reduced the formation of the -ferric uptake regulator protein (-Fur), an inducer of FeSOD expression.

In the present study, overexpression, purification, and characterization of Aeropyrum pernix K1 chaperonin B in E. coli were investigated. The chaperonin -subunit gene (ApCpnB, 1,665 bp ORF) from the hyperthermophilic archaeon A. pernix K1 was amplified by PCR and subcloned into vector pET21a. The constructed pET21a-ApCpnB (6.9 kb) was transformed into E. coli BL21 Codonplus (DE3). The transformant cell successfully expressed ApCpnB, and the expression of ApCpnB (61.2 kDa) was identified through analysis of the fractions by SDS-PAGE (14% gel). The recombinant ApCpnB was purified to higher than 94% by using heat-shock treatment at for 20 min and fast protein liquid chromatography on a HiTrap Q column step. The purified ApCpnB showed ATPase activity and its activity was dependent on temperature. In the presence of ATP, ApCpnB effectively protected citrate synthase (CS) and alcohol dehydrogenase (ADH) from thermal aggregation and inactivation at and , respectively. Specifically, the activity of malate dehydrogenase (MDH) at was greatly stabilized by the addition of ApCpnB and ATP. Coexpression of pro-carboxypeptidase B (pro-CPB) and ApCpnB in E. coli BL21 Codonplus (DE3) had a marked effect on the yield of pro-CPB as a soluble and active form, speculating that ApCpnB facilitates the correct folding of pro-CPB. These results suggest that ApCpnB has both foldase and holdase activities and can be used as a powerful molecular machinery for the production of recombinant proteins as soluble and active forms in E. coli.

Escherichia coli (E. coli) heat-labile enterotoxin B subunit (LTB) was regarded as one of the most powerful mucosal immunoadjuvants eliciting strong immunoresponse to coadministered antigens. In the research, the high-level secretory expression of functional LTB was achieved in P. pastoris through high-density fermentation in a 5-1 fermentor. Meanwhile, the protein was expressed in E. coli by the way of inclusion body, although the gene was cloned from E. coli. Some positive yeast and E. coli transformants were obtained respectively by a series of screenings and identifications. Fusion proteins LTB-6His could be secreted into the supernatant of the medium after the recombinant P. pastoris was induced by 0.5% (v/v) methanol at , whereas E. coli transformants expressed target protein in inclusion body after being induced by 1 mM IPTG at . The expression level increased dramatically to 250-300 mg/l supernatant of fermentation in the former and 80-100 mg/l in the latter. The LTB-6His were purified to 95% purity by affinity chromatography and characterized by SDS-PAGE and Western blot. Adjuvant activity of target protein was analyzed by binding ability with GMI gangliosides. The MW of LTB-6His expressed in P. pastoris was greater than that in E. coli, which was equal to the expected 11 kDa, possibly resulted from glycosylation by P. pastoris that would enhance the immunogenicity of co-administered antigens. These data demonstrated that P. pastoris producing heterologous LTB has significant advantages in higher expression level and in adjuvant activity compared with the homologous E. coli system.

SDS-PAGE of extracted surface-associated proteins of Lactobacillus rhamnosus strains E/N, Oxy, and Pen, was performed. The obtained protein patterns allowed differentiation of the examined strains, which was not accomplished by the commonly used RAPD genotypic method. The differentiation by the SDS-PAGE method proved to be a useful tool for strain-specific identification, which was further confirmed by 2DE analysis. Therefore, it can be used as an alternative or complementary method for both conventional and genotypic identification procedures, especially when closely related lactobacilli isolates are identified.

The Val289 residue in the -amylase of Bacillus amyloliquefaciens, which is equivalent to the Ala289 and Val286 residues in the -amylases of B. stearothermophilus and B. licheniformis, respectively, was studied by site-directed mutagenesis. This residue was substituted with 10 different amino acids by random substitution of the Val codon. In these mutant -amylases, Val289 was substituted with Ile, Tyr, Phe, Leu, Gly, Pro, Ser, Arg, Glu, and Asp. Compared with the wild-type -amylase, the mutant -amylase Val289Ile showed 20% more hydrolytic activity, whereas Val289Phe and Val289Leu showed 50% lesser activity. On the other hand, the mutant -amylases Val289Gly, Val289Tyr, Val289Ser, and Val289Pro showed less than 15% activity. The substitution of Val289 with Arg, Asp, or Glu resulted in complete loss of the -amylase activity. Interestingly, the mutant -amylase Val289Tyr had acquired a transglycosylation activity, which resulted in the change of product profile of the reaction, giving a longer oligosaccharide.

A quantitative detection method for Salmonella in seafood was developed using a SYBR Green-based real-time PCR assay. The assay was developed using pure Salmonella DNA at different dilution levels [i.e., 1,000 to 2 genome equivalents (GE)]. The sensitivity of the real-time assay for Salmonella in seeded seafood samples was determined, and the minimum detection level was 20 CFU/g, whereas a detection level of 2 CFU/ml was obtained for pure culture in water with an efficiency of . The real-time assay was evaluated in repeated experiments with seeded seafood samples and the regression coefficient () values were calculated. The performance of the real-time assay was further assessed with naturally contaminated seafood samples, where 4 out of 9 seafood samples tested positive for Salmonella and harbored cells <100 GE/g, which were not detected by direct plating on Salmonella Chromagar media. Thus, the method developed here will be useful for the rapid quantification of Salmonella in seafood, as the assay can be completed within 2-3 h. In addition, with the ability to detect a low number of Salmonella cells in seafood, this proposed method can be used to generate quantitative data on Salmonella in seafood, facilitating the implementation of control measures for Salmonella contamination in seafood at harvest and post-harvest levels.

Archaebacteria Sulfolobus acidocaldarius contains the highly thermophilic cytochrome P450 enzyme (CYP119). CYP119 possesses stable enzymatic activity at up to . However, this enzyme is still considered as an orphan P450 without known physiological function with endogenous or xenobiotic substrates. We characterized the regioselectivity of lauric acid by CYP119 using the auxiliary redox partner proteins putidaredoxin (Pd) and putidaredoxin reductase (PdR). Purified CYP119 protein showed a tight binding affinity to lauric acid () and dominantly hydroxylated () position of lauric acid. We determined the steady-state kinetic parameters; was 10.8 and , was 12 . The increased ratio to -hydroxylated production of lauric acid catalyzed by CYP119 was observed with increase in the reaction temperature. These studies suggested that the regioselectivity of CYP119 provide the critical clue for the physiological enzyme function in this thermophilic archaebacteria. In addition, regioselectivity control of CYP119 without altering its thermostability can lead to the development of novel CYP119-based catalysts through protein engineering.

A genomic DNA fragment encoding a putative maltooligosyltrehalose synthase (NfMTS) for trehalose biosynthesis was cloned by the degenerate primer-PCR from cyanobacterium Nostoc flagelliforme. The ORF of NfMTS was 2,799 bp in length and encoded 933 amino acid residues constituting a 106.6 kDa protein. The deduced amino acid sequence of NfMTS contained 4 regions highly conserved for MTSs. By expression of NfMTS in E. coli, it was demonstrated that the recombinant protein catalyzed the conversion of maltohexaose to maltooligosyl trehalose. The of the recombinant enzyme for maltohexaose was 1.87 mM and the optimal temperature and pH of the recombinant enzyme was at and 7.0, respectively. The expression of MTS of N. flagelliforme was upregulated, and both trehalose and sucrose contents increased significantly in N. flagelliforme during drought stress. However, trehalose accumulated in small quantities (about 0.36 mg/g DW), whereas sucrose accumulated in high quantities (about 0.90 mg/g DW), indicating both trehalose and sucrose were involved in dehydration stress response in N. flagelliforme and sucrose might act as a chemical chaperone rather than trehalose did during dehydration stress.

Three hundred and seventy-four rhizobacteria were isolated from the rhizosphere soil (RS) or rhizoplane (RP) of Echinochloa crus-galli, Carex leiorhyncha, Commelina communis, Persicaria lapathifolia, Carex kobomugi, and Equisetum arvense, grown in contaminated soil with petroleum and heavy metals. The isolates were screened for plant growth-promoting trait (PGPT), including indole acetic acid (IAA) productivity, 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity, and siderophore(s) synthesis ability. IAA production was detected in 86 isolates (23.0%), ACC deaminase activity in 168 isolates (44.9%), and siderophore(s) synthesis in 213 isolates (57.0%). Among the rhizobacteria showing PGPT, 162 isolates had multiple traits showing more than two types of PGPT. The PGPT-possesing rhizobacteria were more abundant in the RP (82%) samples than the RS (75%). There was a negative correlation (-0.656, p<0.05) between the IAA producers and the ACC deaminase producers. Clustering analysis by principal component analysis showed that RP was the most important factor influencing the ecological distribution and physiological characterization of PGPT-possesing rhizobacteria.

Azotobacter chroococcum H23 (CECT 4435), Azotobacter vinelandii UWD, and Azotobacter vinelandii (ATCC 12837), members of the family Pseudomonadaceae, were used to evaluate their capacity to grow and accumulate polyhydroxyalkanoates (PHAs) using two-phase olive mill wastewater (TPOMW, alpeorujo) diluted at different concentrations as the sole carbon source. The PHAs amounts (g/l) increased clearly when the TPOMW samples were previously digested under anaerobic conditions. The MNR analysis demonstrated that the bacterial strains formed only homopolymers containing -hydroxybutyrate, either when grown in diluted TPOMW medium or diluted anaerobically digested TPOMW medium. COD values of the diluted anaerobically digested waste were measured before and after the aerobic PHA-storing phase, and a clear reduction (72%) was recorded after 72 h of incubation. The results obtained in this study suggest the perspectives for using these bacterial strains to produce PHAs from TPOMW, and in parallel, contribute efficiently to the bioremediation of this waste. This fact seems essential if bioplastics are to become competitive products.

An atrazine-degrading bacterium, strain KU001, was obtained from a sugarcane field at the Cane and Sugar Research and Development Center at the Kasetsart University, Kamphaeng Saen Campus, Thailand. Strain KU001 had a rod-to-coccus morphological cycle during growth. Biolog carbon source analysis indicated that the isolated bacterium was Arthrobacter histidinolovorans. Sequence analysis of the PCR product indicated that the 16S rRNA gene in strain KU001 was 99% identical to the same region in Arthrobacter sp. The atrazine degradation pathway in strain KU001 consisted of the catabolic genes trzN, atzB, and atzC. Strain KU001 was able to use atrazine as a sole nitrogen source for growth, and surprisingly, atrazine degradation was not inhibited in cells grown on ammonium, nitrate, or urea, as compared with cells cultivated on growth-limiting nitrogen sources. During the atrazine degradation process, the supplementation of nitrate completely inhibited atrazine degradation activity in strain KU001, whereas ammonium and urea had no effect on atrazine degradation activity. The addition of strain KU001 to sterile or nonsterile soils resulted in the disappearance of atrazine at a rate that was 4- to 5-fold more than that achieved by the indigenous microbial community. The addition of citrate to soils resulted in enhanced atrazine degradation, where 80% of atrazine disappeared within one day following nutrient supplementation.

The effluent from anaerobic digestion contains organic nitrogen and phosphorus, which are both required for growth of Spirulina platensis. Effluent (20%) from the upflow anaerobic sludge blanket (UASB) from a pig farm, supplemented with 4.5 g/l sodium bicarbonate () and 0.2 g/l urea fertilizer (46:0:0, N:P:K), was found to be not only a suitable medium for the growth of Spirulina platensis but also a low-cost alternative. Cost calculation showed that this medium is 4.4 times cheaper than modifized Zarrouk's medium. The average productivities of a semi-continuous culture grown under outdoor conditions in a 6-1 scale and a 100-1 pilot scale were 19.9 and 12 , respectively. In addition, the biomass of organisms grown in UASB effluent contained approximately 57.9% protein, 1.12% -linolenic acid, and 19.5% phycocyanin. The average rates of bicarbonate, total nitrogen, and phosphorus removal were 380 mg/l/d, 34 mg/l/d, and 4 mg/l/d, respectively.

The viability of low-temperature sulfate reduction with hydrogen as electron donor was studied with a bench-scale gas-lift bioreactor (GLB) operated at . Prior to the GLB experiment, the temperature range of sulfate reduction of the inoculum was assayed. The results of the temperature gradient assay indicated that the inoculum was a psychrotolerant mesophilic enrichment culture that had an optimal temperature for sulfate reduction of , and minimum and maximum temperatures of and , respectively. In the GLB experiment at , a sulfate reduction rate of 500-600 mg , corresponding to a specific activity of 173 mg , was obtained. The electron flow from the consumed -gas to sulfate reduction varied between 27% and 52%, whereas the electron flow to acetate production decreased steadily from 15% to 5%. No methane was produced. Acetate was produced from and by homoacetogenic bacteria. Acetate supported the growth of some heterotrophic sulfate-reducing bacteria. The sulfate reduction rate in the GLB was limited by the slow biomass growth rate at and low biomass retention in the reactor. Nevertheless, this study demonstrated the potential sulfate reduction rate of psychrotolerant sulfate-reducing mesophiles at suboptimal temperature.

Biosynthetic studies on brasiliamides, potently convulsive and bacteriostatic compounds from an endophytic Penicillium brasilianum isolated from Melia azedarach (Meliaceae), confirms their phenylpropanoid origin, which is very uncommon in fungi. Feeding experiments with []-phenylalanine indicated the incorporation of two units of this amino acid on brasiliamide structures. The first step in the phenylpropanoid pathway to those compounds was evaluated through enzymatic bioassays and confirmed the phenylalanine ammonia-lyase (PAL) participation. The metabolism of phenylalanine in this fungus is discussed.

A survey of the endangered orchid Orchis militaris populations was carried out in north-eastern Italy. The occurrence of fungal root endophytes was investigated by light and electron microscopies and molecular techniques. Two main sites of presence were individuated in the Euganean Hills, differing as to the percentage of flowering individuals and of capsules completing maturity. Fluorescence microscopy revealed an intracellular cortical colonization by hyphal pelotons. Two ITS PCR products co-amplified. Sequencing revealed for the former an identity and a high similarity (99%) with a Tulasnellaceae (Basidiomycota) fungus found within tissues of the same host in independent studies in Hungary and Estonia, suggesting an interesting case of tight specificity throughout the Eurosiberian home range. The second amplicon had 99% similarity with Tetracladium species (Ascomycota) recently demonstrated as potential endophytes. TEM revealed two different hyphal structures. Double fungal colonization appears to occur in Orchis militaris and the possible requirement of a specific fungal partner throws light on the causes of this plant's rarity and threatened status.

We investigated the prevalence and the molecular characteristics of vancomycin-intermediate Staphylococcus aureus (VISA) among methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from clinical samples at tertiary or general hospitals participating in a nationwide surveillance program for VISA and vancomycin-resistant Staphylococcus aureus (VRSA) in Korea during an 8-week period in each year from 2001 to 2006. Of 41,639 MRSAs isolated, 37,856 were screened and 169 grew on brain heart infusion agar supplemented with 4 vancomycin. A vancomycin MIC of 4 was confirmed for 33 VISA isolates of the 169 isolates. Eighteen of the 33 isolates were classified as hetero-VISA (hVISA) by the population analysis profile (PAP) method. All VISA isolates were susceptible to linezolid, tigecycline, and quinupristin-dalfopristin. Most VISA isolates (MIC 4 ) showed a PFGE C pattern with sec, seg, and sei enterotoxin genes, including ST5-SCCmec type II, or a PFGE A pattern with sea, including ST239-SCCmec type III.

Evaluation of the primary etiologic agents that cause aseptic meningitis outbreaks may provide valuable information regarding the prevention and management of aseptic meningitis. In Korea, an outbreak of aseptic meningitis caused by echovirus type 30 (E30) occurred from May to October in 2008. In order to determine the etiologic agent, CSF and/or stool specimens from 140 children hospitalized for aseptic meningitis at Soonchunhyang University Cheonan Hospital between June and October of 2008 were tested for virus isolation and identification. E30 accounted for 61.7% (37 cases) and echovirus 6 accounted for 21.7% (13 cases) of all the human enteroviruses (HEVs) isolates (60 cases in total). For the molecular characterization of the isolates, the VP1 gene sequence of 18 Korean E30 isolates was compared pairwise using the MegAlign with 34 reference strains from the GenBank database. The pairwise comparison of the nucleotide sequences of the VP1 genes demonstrated that the sequences of the Korean strains differed from those of lineage groups A, B, C, D, E, F, and G. Reconstruction of the phylogenetic tree based on the complete VP1 nucleotide sequences resulted in a monophyletic tree, with eight clustered lineage groups. All Korean isolates were segregated from other lineage groups, thus suggesting that the Korean strains were a distinct lineage of E30, and a probable cause of this outbreak. This manuscript is the first report, to the best of our knowledge, of the molecular characteristics of E30 strains associated with an aseptic meningitis outbreak in Korea, and their respective phylogenetic relationships.